CN107889744A - The tissue culture and rapid propagation method that a kind of spun gold is seized - Google Patents
The tissue culture and rapid propagation method that a kind of spun gold is seized Download PDFInfo
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- CN107889744A CN107889744A CN201711123613.4A CN201711123613A CN107889744A CN 107889744 A CN107889744 A CN 107889744A CN 201711123613 A CN201711123613 A CN 201711123613A CN 107889744 A CN107889744 A CN 107889744A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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Abstract
The invention provides a kind of spun gold to seize tissue culture and rapid propagation method, is related to plant tissue culture technical field.A kind of spun gold seizes tissue culture medium (TCM)：Initial culture base, proliferated culture medium and root media, primary and proliferated culture medium are Qj culture mediums：N6+6 BA 1.25mg/L+IBA 0.07mg/L+ agar 6.5g/L+ sucrose 30g/L, root media selection Qg culture mediums：1/2MS+6 BA0.01mg/L+IBA 0.5mg/L+ agar 6.5g/L+ sucrose 15g/L；Also include a kind of tissue culture and rapid propagation method using above-mentioned culture medium, including：1) aseptic seedling obtains, and chooses shoot, length processing, is cleaned, sterilized, be degerming, cutting, being inoculated in Qj culture mediums, cultivating to obtain aseptic seedling；2) Multiplying culture, aseptic seedling length processing, is forwarded to culture medium Qj, cultivates to obtain Regenerated plant；3) culture of rootage, Regenerated plant processing are followed by, in Qg culture mediums, cultivating to obtain rooted seedling；4) hardening and transplanting, rooted seedling are transplanted hole tray after moving to greenhouse hardening, transplanted after culture.Effective multiplication rate with culture of rootage is high, and planting percent is high, beneficial to the beneficial effect of factorial praluction.
Description
Technical field
The present invention provides a kind of spun gold and seizes tissue culture and rapid propagation method, is related to field of plant tissue culture technique.
Background technology
Spun gold Chinese catalpa (Catalpa bungei ' jinsiqiu '), Shu Zizang sections (Bignoniaceae) Chinese catalpa category (catalpa)
Tall and big deciduous tree, it is a particularly important high-grade timber-used local varieties in Chinese catalpa (Catalpa bungei), because of sheet material line
There is Herba Anoectochili roxburghii in road and gain the name.Think that spun gold Chinese catalpa is because its variation features is obvious, and with certain adaptation zone, Zhang Junpei etc.
A geographical variety (Zhang Junpei etc., 2008) for Chinese catalpa.Not only material is tough and tensile for spun gold Chinese catalpa, decorative pattern is attractive in appearance, and it has trunk
The advantages that logical straight, strong adaptability, fast speed of growth, it is the preferred seeds of the combination of forestal and agricultural crops, afforestation and afforestation.Gold at present
Silk Chinese catalpa mainly carries out expanding numerous production by means such as root segment cuttage and graftings, and these technological means are short both limited by fertile material
Lack, it is cumbersome, cause spun gold Chinese catalpa reproductive efficiency low, can not solve the shortage of spun gold Chinese catalpa afforestation nursery stock, spun gold Chinese catalpa timber for should not
The social demand problem such as seek.Therefore, it is quickly to improve spun gold Chinese catalpa nursery stock yield urgently to establish spun gold Chinese catalpa high efficiency breeding technology system
The core problem of solution.
Tissue cultures are a kind of efficient asexual reproduction methods.Zhai Xiaoqiao, model Guoqiang etc. (2002) is with spun gold Chinese catalpa children's tender stem
Section thinks that basal medium WPM is better than MS, in culture medium WPM+IBA0.3mg/L+BA5mg/ when carrying out bud induction for explant
Bud induction rate on L is 90%, and induced bud block number is up to 9；1/2MS is better than MS during culture of rootage, in culture medium 1/2MS+
On NAA0.5mg/L, rooting rate is up to 100%.By the end of current, only more than two articles report spun gold Chinese catalpa correlation tissue culture
Research.The research of above-mentioned spun gold Chinese catalpa tissue-culturing rapid propagation only compares to two kinds of minimal mediums, and without embodiment to hormone
The screening of species and proportioning, therefore, the optimal rapid propagation system of spun gold Chinese catalpa is indefinite, and the spun gold Chinese catalpa that structure can be used for production to promote is high
Effect fast breeding technique system still has many technical barriers to need to solve.
Spun gold Chinese catalpa sibling species, kind tissue culture rapid propagation system research in, have common Chinese catalpa method for tissue culture relevant report.
Wherein, Chinese invention patent application CN104054580 A disclose a kind of method for tissue culture for improving Chinese catalpa growth coefficient, bag
Include the foundation of following steps (1) Aseptic culture material；(2) induction of axillary bud；(3) propagation of axillary bud；(4) culture of rootage of sprout；
(5) five steps, the optimal axillary bud deriving formula of the patent application such as hardening and transplanting are MS+BA 0.8mg/L+IBA 0.l mg/
L；Optimal shoot proliferation formula is MS+BA l.5 mg/L+ZT 0.lmg/L+IAA 0.2mg/L；Highest growth coefficient is 15, is led to
Although the method for crossing bottle seedling base portion callus subculture together obtains higher growth coefficient, cannot be directly used to take root, in addition its
Propagation is mainly realized by axillary bud deriving and the step of shoot proliferation two, cumbersome, is unfavorable for promoting in production.Chinese invention is special
Profit application CN104719136A discloses a kind of Chinese catalpa tissue-cultured seedling culture of rootage method, comprises the following steps：(1) Chinese catalpa is sterile
The acquisition of seedling；(2) subculture of aseptic seedling；(3) Chinese catalpa tissue-cultured seedling culture of rootage；(4) four steps such as hardening and transplanting, are applied to
Factorial praluction (height of seedling 2.5-4cm), it is 3-3.5 that it, which effectively expands numerous coefficient, and culture medium is that 6-BA and TDZ is added in MS, is taken root
Cultivate addition IBA 0.5mg/L, rooting rate 96% in the culture medium halved for MS whole elements.Though this method simplifies operation,
But 3-3.5 effective multiplication factor is substantially relatively low, such as carry out the numerous production of batch production expansion using this method, input-output ratio compared with
Small, economic indicator is undesirable.In addition, existing common Chinese catalpa tissue culture method directly is used for into spun gold Chinese catalpa tissue culture, its fast numerous effect is not
It is preferable.
Therefore, culture medium, hormone kind and the proportioning suitable for spun gold Chinese catalpa tissue culture are explored, builds the spun gold of high value-added efficiency
Chinese catalpa rapid propagation system is significant to promoting spun gold Chinese catalpa nursery stock factorial praluction.
The content of the invention
The invention provides a kind of effectively appreciation rate height, the high spun gold of planting percent seizes tissue culture medium (TCM) and tissue-culturing rapid propagation side
Method, solve spun gold in the prior art and seize that tissue culture input-output ratio is small, the undesirable technical problem of fast numerous effect.
To achieve these goals, the present invention uses following technical scheme：
The tissue culture medium (TCM) that a kind of spun gold is seized, including Initial culture base, proliferated culture medium and root media, the primary
Culture medium and the proliferated culture medium are Qj culture mediums, and the root media is Qg culture mediums, and the Qj culture mediums are N6+6-
BA 1.25mg/L+IBA 0.07mg/L+ agar 6.5g/L+ sucrose 30g/L, the Qg culture mediums are 1/2MS+6-BA
0.01mg/L+IBA 0.5mg/L+ agar 6.5g/L+ sucrose 15g/L.
As further preferably, the pH values of the Qj culture mediums is 5.8, the PH values of the Qg culture mediums are 5.8.
A kind of spun gold seizes tissue culture and rapid propagation method, comprises the following steps：
1) spun gold Chinese catalpa aseptic seedling obtains
Take spun gold Chinese catalpa grafting to sprout new buds then in 4-6 months, remove blade, petiole, the tip of a branch and lignifying base portion, cut
Into the long stem segment with axillary bud of 3-5cm, stem with bud is soaked into 10min with saturation detergent water, is placed in after gently brushing surface with banister brush
1h is rinsed under flowing water；On superclean bench, with 75% ethanol postincubation 30s, aseptic water washing 2 times, then with 10% sodium hypochlorite
Solution handles 3min, aseptic water washing 7 times, is inoculated in after being cut into 1.5cm stem with bud in culture medium Qj, and every bottle of inoculation 1 is entered
Row culture, culture obtain spun gold Chinese catalpa aseptic seedling；
2) spun gold Chinese catalpa aseptic seedling squamous subculture
25-28d, long 4-6cm will be cultivated, the base portion that the healthy and strong spun gold Chinese catalpa aseptic seedling of leaf dark green is cut into 0.5-1cm is grown thickly
Bud, it is forwarded in culture medium Qj, every bottle is inoculated with 2, cultivates the spun gold Chinese catalpa tissue-cultured seedling that must be used for taking root or continue subculture；
3) spun gold Chinese catalpa tissue-cultured seedling culture of rootage
Highly it is more than 1cm spun gold Chinese catalpa tissue-cultured seedling in selecting step (2) squamous subculture, retains two blades topmost, other
Blade is then wiped out along petiole base patch cane, is forwarded in root media Qg, and every bottle is inoculated with 2, cultivates to obtain the life of spun gold Chinese catalpa
Root bottle seedling；
4) hardening and transplanting
Spun gold Chinese catalpa rooted seedling band bottle is moved into greenhouse hardening, in 25-28 DEG C of temperature, humidity 80%, fitted under the conditions of natural light
3d is answered, then uncaps and adapts to 3d, cleans culture medium on root system afterwards, transplanting extremely contains turf：Vermiculite is 1:In the hole tray of 1 matrix, treat
After growing young leaves, routinely management continues to cultivate 30d in greenhouse, you can transplanting to crop field.
As further preferred, the condition of culture of step 1)：Light-Dark alternate culture, illumination 14h, intensity of illumination
3000lx, 25 DEG C, dark 10h of temperature, 18 DEG C of temperature, cultivate 25-28d.
As further preferred, the condition of culture of step 2)：Light-Dark alternate culture, illumination 14h, intensity of illumination
3000lx, 25 DEG C, dark 10h of temperature, 18 DEG C of temperature, cultivate 25-28d.
As further preferred, the condition of culture of step 3)：Light-Dark alternate culture, illumination 14h, intensity of illumination
3000lx, 25 DEG C, dark 10h of temperature, 18 DEG C of temperature, cultivate 25-28d.
As further preferably, the base portion Multiple Buds after being cut described in step 2) at least contain 1 axillary bud stem section.
Beneficial effects of the present invention：Without vitrification phenomenon in breeding, Multiplying culture coefficient during Initial culture is reachable
15.5, up to 15.5, the Effective multiplication rate of culture of rootage is high for growth coefficient during Multiplying culture, growth coefficient can 8.1, rooting rate reaches
97.2%, bar number 4.6 of averagely taking root, survival rate is high during acclimatization and transplantses, and emergence rate is up to 95%, beneficial to factorial praluction.
Brief description of the drawings
Fig. 1 is that spun gold seizes Multiplying culture schematic diagram；
Fig. 2 is that spun gold seizes culture of rootage schematic diagram.
Specific embodiment
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clear, complete
Site preparation describes, it is clear that described embodiment is only part of the embodiment of the present invention, rather than whole embodiments.It is based on
Embodiment in the present invention, those of ordinary skill in the art are obtained every other under the premise of creative work is not made
Embodiment, belong to the scope of protection of the invention.
Embodiment 1
1) spun gold Chinese catalpa aseptic seedling obtains
Take spun gold Chinese catalpa grafting to sprout new buds then in 4-6 months, remove blade, petiole, the tip of a branch and lignifying base portion, cut
Into the long stem segment with axillary bud of 3-5cm, stem with bud is soaked into 10min with saturation detergent water, is placed in after gently brushing surface with banister brush
1h is rinsed under flowing water；On superclean bench, with 75% ethanol postincubation 30s, aseptic water washing 2 times, then with 10% sodium hypochlorite
Solution handles 3min, aseptic water washing 7 times, is inoculated in Initial culture base after being cut into 1.5cm stem with bud, Initial culture base is
Qj culture mediums, mg/L+IBA 0.07mg/L+ agar 6.5g/L+ the sucrose 30g/L, PH 5.8 of N6+6-BA 1.25, every bottle of inoculation
1, condition of culture：Light-Dark alternate culture, illumination 14h, intensity of illumination 3000lx, 25 DEG C, dark 10h of temperature, temperature 18
DEG C, 25-28d is cultivated, obtains spun gold Chinese catalpa aseptic seedling.
2) spun gold Chinese catalpa aseptic seedling squamous subculture
25-28d, long 4-6cm will be cultivated, the healthy and strong spun gold Chinese catalpa aseptic seedling of leaf dark green is cut into 0.5-1cm and at least contains 1 armpit
Leaf stem section and base portion Multiple Buds, are forwarded to proliferated culture medium together, and proliferated culture medium is Qj culture mediums：N6+6-BA 1.25mg/L
+ IBA 0.07mg/L+ agar 6.5g/L+ sucrose 30g/L, PH 5.8, every bottle is inoculated with 2.Condition of culture：Light-Dark replaces
Culture, illumination 14h, intensity of illumination 3000lx, 25 DEG C, dark 10h of temperature, 18 DEG C of temperature, 25-28d is cultivated, must can be used to take root
Or continue the spun gold Chinese catalpa tissue-cultured seedling of subculture.
3) spun gold Chinese catalpa tissue-cultured seedling culture of rootage
Highly it is more than 1cm spun gold Chinese catalpa tissue-cultured seedling in selecting step (2) squamous subculture, retains two blades topmost, other
Blade is then wiped out along petiole base patch cane, is forwarded to root media Qg, Qg： 1/2MS+6-BA 0.01mg/L+
IBA0.5mg/L+ agar 6.5g/L+ sucrose 15g/L, PH 5.8, every bottle is inoculated with 2.Condition of culture：Light-Dark is alternately trained
Support, illumination 14h, intensity of illumination 3000lx, 25 DEG C, dark 10h of temperature, 18 DEG C of temperature, cultivate 25-28d, obtain the life of spun gold Chinese catalpa
Root bottle seedling, rooting rate 97.2%.
4) hardening and transplanting
Spun gold Chinese catalpa rooted seedling band bottle is moved into greenhouse hardening, in 25-28 DEG C of temperature, humidity 80%, fitted under the conditions of natural light
3d is answered, then uncaps and adapts to 3d, cleans culture medium on root system afterwards, transplanting extremely contains turf：Vermiculite is 1:In the hole tray of 1 matrix, treat
After growing young leaves, routinely management continues to cultivate 30d in greenhouse, you can transplanting is to crop field, and emergence rate is up to more than 95%.
Culture medium choice experiment
1. the screening of Qj culture mediums in the acquisition of spun gold Chinese catalpa aseptic seedling, spun gold Chinese catalpa aseptic seedling squamous subculture
According to hormone concentration of the data primary election suitable for spun gold Chinese catalpa tissue culture propagation has been announced, select to add not in WPM culture mediums
With concentration 6-BA and NAA, while agar 6.5g/L is added, sucrose 30g/L, regulation PH ≈ 5.8.Each processing is inoculated with not during inoculation
Less than 30 bottles, culture 25-28d investigation growing states, 1 is specifically shown in Table.
6-BA the and NAA comparative experiments of table 1
Wherein value-added coefficient 1 refers to the multiplied ratio of base portion Multiple Buds subculture together；Value-added coefficient 2 refers to height and is more than 1cm buds
The multiplied ratio of (can be directly used for culture of rootage).
It can be seen that 6-BA 1.3mg/L, NAA 0.13mg/L break up beneficial to spun gold Chinese catalpa, but generally existing is slow-growing, leaf
The problems such as color turns to be yellow.Suitable minimal medium can not only promote the robust growth of spun gold Chinese catalpa, also have certain promotion to make to propagation
With.Therefore, it is further optimization, 6-BA 1.3mg/L, NAA 0.13mg/L agar 6.5g/ is added in different minimal mediums
L, sucrose 30g/L, regulation PH ≈ 5.8, culture 25-28d investigation growing states, screening are suitable to the fast numerous basic culture of spun gold Chinese catalpa
Base, concrete outcome are shown in Table 2.
The different culture media of table 2 is tested
| Numbering | Culture medium | Value-added coefficient 1 | Growth coefficient 2 | Description |
| 1 | 1/2MS | 6.5 | 2.3 | Yellow leaf, slow-growing, growth coefficient is low |
| 2 | MS | 9.2 | 4.1 | Leaf dark green is carried out, robust plant, and growth coefficient is high |
| 3 | N6 | 9.5 | 4.3 | Robust plant, leaf green, growth coefficient highest |
| 4 | WPM | 6.3 | 1.7 | Leaf green, plant strain growth is slow, and growth coefficient is low |
| 5 | DKW | 5.5 | 1.9 | Yellow leaf, plant strain growth is slow, and value-added coefficient is low |
As a result show, N6 culture mediums are more suitable for spun gold Chinese catalpa tissue culture than other minimal mediums.Further optimization, cultivated in N6
The auxin and 6-BA 1.3mg/L of variety classes concentration are added in base, agar 6.5g/L, sucrose 30g/L, adjusts PH ≈ 5.8,
25-28d investigation growing states are cultivated, concrete outcome is shown in Table 3.
Table 3 IBA, NAA, IAA hormone choice experiment
| Numbering | Auxin | Value-added coefficient 1 | Growth coefficient 2 | Description |
| 1 | NAA 0.13mg/L | 9.3 | 3.7 | Value-added effect is general, and it is few to can be directly used for culture of rootage quantity |
| 2 | IBA0.1mg/L | 11.5 | 4.3 | Value-added effect is general, and it is on the low side to can be directly used for culture of rootage quantity |
| 3 | IBA0.07mg/L | 13.6 | 6.1 | Value-added coefficient is high, and it is more to can be directly used for culture of rootage quantity |
| 4 | IBA0.05mg/L | 12.8 | 4.5 | Value-added effect is general, and it is on the low side to can be directly used for culture of rootage quantity |
| 5 | IAA0.13mg/L | 3.3 | 1.2 | Growth coefficient is low, and it is few to can be directly used for culture of rootage quantity |
| 6 | IAA0.1mg/L | 4.2 | 1.4 | Growth coefficient is low, and it is few to can be directly used for culture of rootage quantity |
| 7 | IAA0.07mg/L | 4.3 | 1.1 | Growth coefficient is low, and it is few to can be directly used for culture of rootage quantity |
As a result show：IBA is more suitable for spun gold Chinese catalpa tissue-culturing rapid propagation than NAA, IAA, adds IBA 0.07mg/L, the increment of spun gold Chinese catalpa
Coefficient is high, and growth is fast, and it is more to can be directly used for culture of rootage tissue-cultured seedling quantity.For further optimization, screening is suitable to spun gold Chinese catalpa tissue culture
Hormone kind and proportioning, agar 6.5g/L is being with the addition of, added in sucrose 30g/L, PH ≈ 5.8 N6 culture mediums following different sharp
Element, culture 25-28d investigation growing states, experimental result are shown in Table 4.
Table 4 is different to cross hormone kind choice experiment
| Numbering | Hormone kind | Value-added coefficient 1 | Growth coefficient 2 | Description |
| 1 | 6-BA+IBA | 15.1 | 7.5 | Robust plant, it is more to can be directly used for culture of rootage quantity |
| 2 | 6-BA+TDZ | 8.5 | 5.3 | Robust plant, it is on the low side to can be directly used for culture of rootage quantity |
| 3 | 6-BA+ZT+IAA | 13.5 | 1.8 | Easily there is vitrifying, it is few to can be directly used for culture of rootage quantity |
| 4 | 6-BA+KT+IBA | 3.5 | 1.1 | Easily there is vitrifying, value-added coefficient is low |
| 5 | ZT+IBA | 2.7 | 0.9 | Value-added coefficient is low or even undifferentiated |
| 6 | KT+IBA | 3.5 | 1.5 | Growth coefficient is low or even undifferentiated |
| 7 | KT+TDZ+IBA | 2.5 | 0.7 | Easily there is vitrifying, value-added coefficient is low |
As a result show:6-BA and IBA is used alone just can produce fine cultivation effect to spun gold Chinese catalpa tissue culture, and plant strain growth is good for
Strong, the tissue-cultured seedling quantity that generation can be directly used for culture of rootage is more, and single use 6-BA is more cost-effective, is above pushed away beneficial in production
Extensively.For further optimization 6-BA concentration, in addition IBA 0.07mg/L, agar 6.5g/L, sucrose 30g/L, PH ≈ 5.8 N6 is trained
Support and various concentrations 6-BA is added in base, experimental result is shown in Table 5.
The 6-BA concentration of table 5 selects
| Numbering | 6-BA concentration | Value-added coefficient 1 | Growth coefficient 2 | Description |
| 1 | 1.0mg/L | 6.4 | 2.5 | Robust growth, value-added coefficient are low |
| 2 | 1.2mg/L | 12.2 | 4.3 | Robust growth, growth coefficient are low |
| 3 | 1.25mg/L | 15.5 | 8.1 | 3 holding higher growth efficiency of subculture, no vitrification phenomenon |
| 4 | 1.3mg/L | 15.1 | 6.1 | 3 value-added coefficients of subculture reduce, and vitrifying occurs in 8% bottle seedling |
Due to subculture number increase, hormone high concentration 6-BA after plant interior accumulation, 3 subcultures easily causes vitrifying
The problems such as seedling occurs, and value-added coefficient declines, therefore the culture medium for being preferably suitable to spun gold Chinese catalpa propagation is N6+6-BA 1.25mg/L+
IBA 0.07mg/L, agar 6.5g/L, sucrose 30g/L, regulation PH ≈ 5.8.
2. the screening of Qg culture mediums in step (3)
Added containing agar 6.5g/L, in sucrose 15g/L, PH ≈ 5.8 1/2MS culture mediums various concentrations 6-BA and
IBA, culture 25-28d investigate situation of taking root, are specifically shown in Table 6.
The Qg culture medium choice experiments of table 6
As a result show：On 1/2MS+6-BA0.01mg/L+IBA 0.5mg/L culture mediums, spun gold Chinese catalpa rooting efficiency is best,
Robust plant, rooting rate is up to 97.2%, bar number 4.6 of averagely taking root, and easily survives during acclimatization and transplantses.
Explanation：Halved in screening process using 1/2MS culture mediums for a great number of elements in MS culture mediums；Above-mentioned screening process is only
List the part for producing significant result, and unlisted whole screening processes.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
God and any modification, equivalent substitution, improvement etc. within principle, made, should be included in the scope of the protection.
Claims (7)
- A kind of 1. tissue culture medium (TCM) that spun gold is seized, it is characterised in that including Initial culture base, proliferated culture medium and culture of rootage Base, the Initial culture base and the proliferated culture medium are Qj culture mediums, and the root media is Qg culture mediums, the Qj trainings It is N6+6-BA 1.25mg/L+IBA 0.07mg/L+ agar 6.5g/L+ sucrose 30g/L to support base, and the Qg culture mediums are 1/2MS+ 6-BA 0.01mg/L+IBA 0.5mg/L+ agar 6.5g/L+ sucrose 15g/L.
- 2. the tissue culture medium (TCM) that a kind of spun gold according to claim 1 is seized, it is characterised in that the pH value of the Qj culture mediums For 5.8, the pH value of the Qg culture mediums is 5.8.
- 3. the spun gold for the tissue culture medium (TCM) seized using a kind of spun gold described in claim 1 seizes tissue culture and rapid propagation method, its feature exists In comprising the following steps：1) spun gold Chinese catalpa aseptic seedling obtainsTake spun gold Chinese catalpa grafting to sprout new buds then in 4-6 months, remove blade, petiole, the tip of a branch and lignifying base portion, be cut into 3- The long stem segment with axillary bud of 5cm, stem with bud is soaked into 10min with saturation detergent water, flowing water is placed in after gently brushing surface with banister brush Lower flushing 1h；On superclean bench, with 75% ethanol postincubation 30s, aseptic water washing 2 times, then with 10% liquor natrii hypochloritis 3min is handled, aseptic water washing 7 times, is inoculated in after being cut into 1.5cm stem with bud in culture medium Qj, every bottle of inoculation 1 is trained Support, culture obtains spun gold Chinese catalpa aseptic seedling；2) spun gold Chinese catalpa aseptic seedling squamous subculture25-28d, long 4-6cm will be cultivated, the healthy and strong spun gold Chinese catalpa aseptic seedling of leaf dark green is cut into 0.5-1cm base portion Multiple Buds, turned It is connected in culture medium Qj, every bottle is inoculated with 2, and culture obtains the spun gold Chinese catalpa tissue-cultured seedling for taking root or continuing subculture；3) spun gold Chinese catalpa tissue-cultured seedling culture of rootageHighly it is more than 1cm spun gold Chinese catalpa tissue-cultured seedling in selecting step (2) squamous subculture, retains two blades topmost, other blades Then wipe out, be forwarded in root media Qg along petiole base patch cane, every bottle is inoculated with 2, cultivate spun gold Chinese catalpa takes root bottle Seedling；4) hardening and transplantingSpun gold Chinese catalpa rooted seedling band bottle is moved into greenhouse hardening, in 25-28 DEG C of temperature, humidity 80%, adapts to 3d under the conditions of natural light, Uncap again and adapt to 3d, clean culture medium on root system afterwards, transplanting extremely contains turf：Vermiculite is 1:In the hole tray of 1 matrix, wait to grow newly Ye Hou, routinely management continue to cultivate 30d in greenhouse, you can transplanting to crop field.
- 4. a kind of spun gold according to claim 3 seizes tissue culture and rapid propagation method, it is characterised in that the condition of culture of step 1)： Light-Dark alternate culture, illumination 14h, intensity of illumination 3000lx, 25 DEG C, dark 10h of temperature, 18 DEG C of temperature, cultivate 25- 28d。
- 5. a kind of spun gold according to claim 4 seizes tissue culture and rapid propagation method, it is characterised in that the condition of culture of step 2)： Light-Dark alternate culture, illumination 14h, intensity of illumination 3000lx, 25 DEG C, dark 10h of temperature, 18 DEG C of temperature, cultivate 25- 28d。
- 6. a kind of spun gold according to claim 4 seizes tissue culture and rapid propagation method, it is characterised in that the condition of culture of step 3)： Light-Dark alternate culture, illumination 14h, intensity of illumination 3000lx, 25 DEG C, dark 10h of temperature, 18 DEG C of temperature, cultivate 25- 28d。
- 7. tissue culture and rapid propagation method is seized according to a kind of any described spun golds of claim 3-6, it is characterised in that described in step 2) Cut after base portion Multiple Buds at least contain 1 axillary bud stem section.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108739408A (en) * | 2018-08-01 | 2018-11-06 | 河南省农业科学院 | A kind of method for tissue culture improving the effective shoot differentiation quantity of great Ye spun gold Chinese catalpas |
| CN109329064A (en) * | 2018-11-26 | 2019-02-15 | 山东省林业科学研究院 | A kind of method for tissue culture and special culture media improving Chinese catalpa growth coefficient |
| CN111213584A (en) * | 2020-02-03 | 2020-06-02 | 鲁东大学 | Method for regenerating plants by high-frequency induction of kalopanax septemlobus leaves |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103190344A (en) * | 2013-03-29 | 2013-07-10 | 中国林业科学研究院林业研究所 | Tissue culture method of fargesii |
-
2017
- 2017-11-14 CN CN201711123613.4A patent/CN107889744B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103190344A (en) * | 2013-03-29 | 2013-07-10 | 中国林业科学研究院林业研究所 | Tissue culture method of fargesii |
Non-Patent Citations (2)
| Title |
|---|
| 翟晓巧等: "金丝楸茎段的组织培养及植株再生技术研究", 《河南农业大学学报》 * |
| 范国强等: "金丝楸的离体培养和植株再生", 《植物生理学通讯》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108739408A (en) * | 2018-08-01 | 2018-11-06 | 河南省农业科学院 | A kind of method for tissue culture improving the effective shoot differentiation quantity of great Ye spun gold Chinese catalpas |
| CN108739408B (en) * | 2018-08-01 | 2021-05-18 | 河南省农业科学院 | Tissue culture method for increasing effective young shoot differentiation number of golden mountain ash |
| CN109329064A (en) * | 2018-11-26 | 2019-02-15 | 山东省林业科学研究院 | A kind of method for tissue culture and special culture media improving Chinese catalpa growth coefficient |
| CN111213584A (en) * | 2020-02-03 | 2020-06-02 | 鲁东大学 | Method for regenerating plants by high-frequency induction of kalopanax septemlobus leaves |
| CN111213584B (en) * | 2020-02-03 | 2022-03-29 | 鲁东大学 | Method for regenerating plants by high-frequency induction of kalopanax septemlobus leaves |
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| CN107889744B (en) | 2019-08-13 |
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